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Tailoring butyl methacrylate/methacrylic acid copolymers for the solubilization of membrane proteins: the influence of composition and molecular weight

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    0562629 - ÚMCH 2023 RIV DE eng J - Journal Article
    Janata, Miroslav - Čadová, Eva - Angelisová, Pavla - Charnavets, Tatsiana - Hořejší, Václav - Raus, Vladimír
    Tailoring butyl methacrylate/methacrylic acid copolymers for the solubilization of membrane proteins: the influence of composition and molecular weight.
    Macromolecular Bioscience. Roč. 22, č. 10 (2022), č. článku 2200284. ISSN 1616-5187. E-ISSN 1616-5195
    R&D Projects: GA ČR(CZ) GA19-04047S; GA MŠMT(CZ) LM2018127
    Research Infrastructure: CIISB II - 90127
    Institutional support: RVO:61389013 ; RVO:68378050 ; RVO:86652036
    Keywords : amphiphilic copolymers * isolation * membrane proteins
    OECD category: Polymer science; Biochemistry and molecular biology (UMG-J); Biochemistry and molecular biology (BTO-N)
    Impact factor: 4.6, year: 2022
    Method of publishing: Limited access
    https://onlinelibrary.wiley.com/doi/10.1002/mabi.202200284

    Low-molecular weight (MW) amphiphilic copolymers have been recently introduced as a powerful tool for the detergent-free isolation of cell membrane proteins. Herein, a screening approach is used to identify a new copolymer type for this application. Via a two-step ATRP/acidolysis procedure, a 3 × 3 matrix of well-defined poly[(butyl methacrylate)-co-(methacrylic acid)] copolymers (denoted BMAA) differing in their MW and ratio of hydrophobic (BMA) and hydrophilic (MAA) units is prepared. Subsequently, using the biologically relevant model (T-cell line Jurkat), two compositions of BMAA copolymers are identified that solubilize cell membranes to an extent comparable to the industry standard, styrene-maleic acid copolymer (SMA), while avoiding the potentially problematic phenyl groups. Surprisingly, while only the lowest-MW variant of the BMA/MAA 2:1 composition is effective, all the copolymers of the BMA/MAA 1:1 composition are found to solubilize the model membranes, including the high-MW variant (MW of 14 000). Importantly, the density gradient ultracentrifugation/sodium dodecyl sulfate-polyacrylamide gel electrophoresis/Western blotting experiments reveal that the BMA/MAA 1:1 copolymers disintegrate the Jurkat membranes differently than SMA, as demonstrated by the different distribution patterns of two tested membrane protein markers. This makes the BMAA copolymers a useful tool for studies on membrane microdomains differing in their composition and resistance to membrane-disintegrating polymers.
    Permanent Link: https://hdl.handle.net/11104/0335277

     
     
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